Methods and panels of compounds for characterization of glioblastoma multiforme tumors and cancer stem cells thereof

Abstract

A method of characterizing a glioblastoma multiforme (GBM) stem cell (GSC), comprising culturing the GSC to provide a culture, contacting a first set of aliquots of the culture with individual compounds selected from a panel of compounds, identifying two or more of the selected compounds that cause more than a threshold level of cell death in the first set of aliquots, and characterizing the GSC as suitable for treatment with one or more combinations comprising the two or more identified compounds. A panel of chemical compounds, the compounds selected by a method comprising surgically resecting the tumor, culturing a GSC derived from GBM tissue derived from a GBM tumor, contacting aliquots thereof with individual compounds selected from a panel of compounds, and identifying two or more of the selected compounds that cause more than a threshold level of cell death in the aliquots, thereby identifying the compounds.

What is claimed is: 
     
       1. A method of treating a patient having a glioblastoma multiforme (GBM) tumor comprising a GBM stem cell in order to extend the patient's overall survival following treatment with three or more identified compounds , comprising:
 a) culturing the GBM stem cell to provide a culture; 
 b) contacting a first set of aliquots of the culture with individual compounds selected from a panel of compounds; 
 c) identifying three or more of the selected compounds that cause more than a threshold level of cell death in the first set of aliquots; 
 d) characterizing the patient having the GBM tumor comprising the GBM stem cell as suitable for treatment with one or more combinations comprising the three or more identified compounds; and 
 e) administering one or more combinations comprising the three or more identified compounds to the patient, thereby treating the patient having the GBM tumor 
 wherein the panel comprises at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, or all, of the following compounds: AZD-7762; AZD-8055; BEZ-235; BGT-226; MG-132; NVP-AUY-922; obatoclax; PIK-75; PKI-587; staurosporine; YM155; 10-hydroxycamptothecin; anisomycin; β-peltatin; emetine; gambogic acid; mebendazole; ouabain; peruvoside; phenethyl caffeate; podophyllin acetate; pyrvinium pamoate; and thiram, and 
 wherein the panel further comprises at least one of: auranofin, bortezomib, colchicine, digoxin, disulfiram, methotrexate, pitavastatin, sertraline, valganciclovir, and vincristine sulfate; and wherein the panel further comprises one or more of the following compounds: albendazole, aprepitant, atorvastatin calcium, bleomycin, brompheniramine maleate, captopril, carboplatin, celecoxib, chloroquine, cidofovir, cladribine, clofarabine, clofazimine, clomipramine, clonidine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, docetaxel, doxorubicin, erlotinib, ethacrynic acid, epirubicin, etoposide, fludarabine phosphate, fluorouracil, fluphenazine, fluvastatin, gemcitabine, haloperidol, homoharringtonine, hydroxychloroquine, idarubicin, irinotecan, itraconazole, ketoconazole, lomustine, lovastatin, mefloquine, melphalan, memantine, metformin, methylene blue, minoxidil, mitomycin C, mitoxantrone, mycophenolate mofetil, nelfinavir, nisoldipine, paclitaxel, pindolol, rosuvastatin calcium, simvastatin, sirolimus, sorafenib, sodium tetradecyl sulfate, sunitinib, temozolomide, teniposide, topotecan, trifluoperazine, valproic acid, vinblastine sulfate, vinorelbine and vorinostat, 
 wherein the patient has completed a standard of care therapy comprising radiation and temozolomide (TMZ). 
 
     
     
       2. The method of  claim 1 , wherein the GBM stem cell is derived from a GBM tumor tissue, and wherein the culturing step (a) comprises:
 i) adding one or more dissociation enzymes to the GBM tumor tissue; 
 ii) incubating the GBM tumor tissue to facilitate digestion of the GBM tumor tissue by the one or more enzymes; 
 iii) obtaining a suspension of cells from the digested GBM tumor tissue; and 
 iv) propagating the suspension of cells in stem cell culture medium to provide the culture. 
 
     
     
       3. The method of  claim 2 , wherein the dissociation enzyme is an enzyme mixture with proteolytic and collagenolytic enzyme activity. 
     
     
       4. The method of  claim 2 , wherein the incubating takes place at a temperature range between 35° C. and 39° C. 
     
     
       5. The method of  claim 4 , wherein the incubating takes place at about 37° C. 
     
     
       6. The method of  claim 1 , wherein the threshold level is about 50%. 
     
     
       7. The method of  claim 1 , wherein at least one of the one or more combinations comprises at least one anti-neoplastic/chemotherapeutic compound and at least one additional compound. 
     
     
       8. The method of  claim 1 , wherein step (c) further comprises:
 from the identified three or more selected compounds, further selecting at least two compounds as preferred compounds based on at least one of the following characteristics:
 overall level of cell death caused in the first set of aliquots; 
 known ability to cross the blood-brain barrier in a GBM patient; and 
 known likely side effects to a GBM patient, and 
 
 wherein step (d) further comprises characterizing the GBM stem cell as suitable for treatment with one or more combinations of the preferred compounds.